{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T12:06:56Z","timestamp":1789042016863,"version":"build-2803163510"},"posted":{"date-parts":[[2026,9,7]]},"group-title":"Microbiology","reference-count":60,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T00:00:00Z","timestamp":1788739200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["1R16AI189394-01"],"award-info":[{"award-number":["1R16AI189394-01"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"award":["P20GM121176"],"award-info":[{"award-number":["P20GM121176"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000997","name":"Arnold and Mabel Beckman Foundation","doi-asserted-by":"crossref","award":["Beckman Scholars Program"],"award-info":[{"award-number":["Beckman Scholars Program"]}],"id":[{"id":"10.13039\/100000997","id-type":"DOI","asserted-by":"crossref"}]},{"award":["Major Research Instrumentation award (2017945)"],"award-info":[{"award-number":["Major Research Instrumentation award (2017945)"]}],"id":[{"id":"https:\/\/ror.org\/021nxhr62","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Analytical Resources Core - Colorado State University"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,7]]},"abstract":"<jats:label>2.<\/jats:label>\n                <jats:title>ABSTRACT AND GRAPHICAL ABSTRACT<\/jats:title>\n                <jats:p>\n                  Visceral leishmaniasis (VL) is a neglected tropical disease caused by the protozoan parasites\n                  <jats:italic>Leishmania infantum<\/jats:italic>\n                  (LI) and\n                  <jats:italic>Leishmania donovani<\/jats:italic>\n                  (LD). VL disproportionately affects populations that live in extreme poverty and those affected by climate change. Treatment options are limited, fiscally prohibitive, and toxic to some that receive treatment. In cases where VL remains untreated, death is inevitable. Although disease prevalence is increasing in affluent countries with robust research capabilities, there remains little economic incentive to search for new, more accessible non-toxic treatments, emphasizing the neglected tropical aspect of this disease. Our previous results indicate that sulforaphane (SFN) inhibits VL promastigote and amastigote growth through an unknown mechanism. The current study extends the anti-parasitic effects of SFN establishing SFN\u2019s efficacy against multiple\n                  <jats:italic>Leishmania<\/jats:italic>\n                  parasites including\n                  <jats:italic>L. tropica, L. mexicana, L. panamensis<\/jats:italic>\n                  and\n                  <jats:italic>L. amazonesis<\/jats:italic>\n                  . Moreover, SFN was an effective therapy post-infection as stably infected macrophages treated with SFN (10 \u00b5M for 48 hours) had a significant reduction in the number of infected cells and the number of amastigotes per cell against\n                  <jats:italic>Leishmania<\/jats:italic>\n                  species that cause both VL and cutaneous leishmaniasis (CL). Moreover, SFN led to a reduction of motility within 6 hours of treatment indicating a direct interaction with the parasite. Alkynyl SFN (A-SFN) was subsequently synthesized to determine the specific binding target of SFN. Interestingly, A-SFN was more effective than SFN in decreasing parasite viability and inhibiting motility against VL and CL parasite species at equivalent concentrations. Immunofluorescence analysis utilizing click chemistry indicated A-SFN binds to cytosolic promastigote components and colocalized only minimally with KMP-11. The binding of SFN to KMP-11 could not be confirmed\n                  <jats:italic>in vitro.<\/jats:italic>\n                  In terms of intracellular host effects, SFN treatment increased the induction of nonselective macroautophagy by increased lysosomal acidification as quantified by DALGreen and FITC-dextran staining in infected macrophages. The induction of autophagy occurred simultaneously with the upregulation of expression of cytoprotective, antioxidant and autophagy-related genes via the NRF2 transcription factor. These data provide robust evidence that SFN is an excellent lead compound against\n                  <jats:italic>Leishmania<\/jats:italic>\n                  that initiates multiple innate immune pathways that enable enhanced parasite killing. Future studies will focus on the\n                  <jats:italic>in vivo<\/jats:italic>\n                  efficacy of SFN against CL.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Graphical abstract<\/jats:title>\n                  <jats:fig id=\"ufig1\" position=\"float\" orientation=\"portrait\" fig-type=\"figure\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"749349v1_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:label>3.<\/jats:label>\n                  <jats:title>KEY FINDINGS<\/jats:title>\n                  <jats:list list-type=\"bullet\">\n                    <jats:list-item>\n                      <jats:p>\n                        Sulforaphane (SFN) decreased promastigote viability from six\n                        <jats:italic>Leishmania<\/jats:italic>\n                        species\n                      <\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>SFN is an effective therapy that decreased the number of intracellular VL and CL amastigotes in macrophages post-infection<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>SFN inhibited promastigote motility within 6 hours<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>\n                        SFN increased NRF2-dependent pathway expression and lysosomal acidification in cells infected by\n                        <jats:italic>Leishmania<\/jats:italic>\n                      <\/jats:p>\n                    <\/jats:list-item>\n                  <\/jats:list>\n                <\/jats:sec>","DOI":"10.64898\/2026.09.04.749349","type":"posted-content","created":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T06:05:16Z","timestamp":1788847516000},"source":"Crossref","is-referenced-by-count":0,"title":["Sulforaphane targets\n                  <i>Leishmania<\/i>\n                  promastigotes to inhibit parasite motility and alters host autophagy to suppress parasite replication in macrophages"],"prefix":"10.64898","author":[{"given":"Elenora","family":"Kelley","sequence":"first","affiliation":[{"name":"Department of Biology, Fort Lewis College, Durango, CO, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emmet","family":"Thompson","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, Fort Lewis College, Durango, CO, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahliya","family":"Harding","sequence":"additional","affiliation":[{"name":"Department of Biology, Fort Lewis College, Durango, CO, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katie","family":"Torrey","sequence":"additional","affiliation":[{"name":"Department of Biology, Fort Lewis College, Durango, CO, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai Gurnoe","family":"Brantley","sequence":"additional","affiliation":[{"name":"Department of Health and Exercise Science, Colorado State University, Fort Collins, CO, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunyu","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Drug Design and Pharmacology, University of Copenhagen, DK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anders","family":"Bach","sequence":"additional","affiliation":[{"name":"Department of Drug Design and Pharmacology, University of Copenhagen, DK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenneth A.","family":"Miller","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, Fort Lewis College, Durango, CO, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9277-2475","authenticated-orcid":false,"given":"David J.","family":"Blake","sequence":"additional","affiliation":[{"name":"Department of Biology, Fort Lewis College, Durango, CO, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026091004100726000_2026.09.04.749349v1.1","doi-asserted-by":"publisher","DOI":"10.1007\/s40475-021-00232-7"},{"key":"2026091004100726000_2026.09.04.749349v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ebiom.2023.104440"},{"key":"2026091004100726000_2026.09.04.749349v1.3","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMra2403309"},{"key":"2026091004100726000_2026.09.04.749349v1.4","first-page":"471","article-title":"Global Leishmaniasis surveillance, 2022: assessing trends over the past 10 years","volume":"98","year":"2023","journal-title":"Weekly Epidemiological Record"},{"key":"2026091004100726000_2026.09.04.749349v1.5","doi-asserted-by":"publisher","DOI":"10.1038\/s41572-025-00663-w"},{"key":"2026091004100726000_2026.09.04.749349v1.6","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.4c00778"},{"key":"2026091004100726000_2026.09.04.749349v1.7","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.4c00339"},{"key":"2026091004100726000_2026.09.04.749349v1.8","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms9030578"},{"key":"2026091004100726000_2026.09.04.749349v1.9","doi-asserted-by":"publisher","DOI":"10.1001\/JAMADERMATOL.2018.2133"},{"key":"2026091004100726000_2026.09.04.749349v1.10","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms13112485"},{"key":"2026091004100726000_2026.09.04.749349v1.11","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0014297"},{"key":"2026091004100726000_2026.09.04.749349v1.12","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.20-0107"},{"key":"2026091004100726000_2026.09.04.749349v1.13","doi-asserted-by":"crossref","unstructured":"Jardim A , Funk V , Capriolit RM , Olafson RW . Isolation and structural characterization of the Leishmania donovani kinetoplastid membrane protein-11, a major immunoreactive membrane glycoprotein. Biochem J. 1995.","DOI":"10.1042\/bj3050307"},{"key":"2026091004100726000_2026.09.04.749349v1.14","doi-asserted-by":"publisher","DOI":"10.1128\/EC.00249-08"},{"key":"2026091004100726000_2026.09.04.749349v1.15","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2015.00001"},{"key":"2026091004100726000_2026.09.04.749349v1.16","doi-asserted-by":"publisher","DOI":"10.1590\/S0074-02762010000300018"},{"key":"2026091004100726000_2026.09.04.749349v1.17","doi-asserted-by":"publisher","DOI":"10.1590\/S0074-02761998000200022"},{"key":"2026091004100726000_2026.09.04.749349v1.18","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2249.2008.03741.x"},{"key":"2026091004100726000_2026.09.04.749349v1.19","doi-asserted-by":"publisher","DOI":"10.1128\/iai.62.11.4893-4899.1994"},{"key":"2026091004100726000_2026.09.04.749349v1.20","doi-asserted-by":"publisher","DOI":"10.2307\/3284745"},{"key":"2026091004100726000_2026.09.04.749349v1.21","doi-asserted-by":"publisher","DOI":"10.1038\/s44319-024-00302-7"},{"key":"2026091004100726000_2026.09.04.749349v1.22","first-page":"4632","article-title":"Mechanism of Differential Potencies of Isothiocyanates as Inducers of Anticarcinogenic Phase 2 Enzymes1","volume":"58","year":"1998","journal-title":"Cancer Res"},{"key":"2026091004100726000_2026.09.04.749349v1.23","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmc.2022.117136"},{"key":"2026091004100726000_2026.09.04.749349v1.24","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202508121"},{"key":"2026091004100726000_2026.09.04.749349v1.25","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2023.102248"},{"key":"2026091004100726000_2026.09.04.749349v1.26","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1005595"},{"key":"2026091004100726000_2026.09.04.749349v1.27","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-022-12347-6"},{"key":"2026091004100726000_2026.09.04.749349v1.28","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejmech.2013.03.045"},{"key":"2026091004100726000_2026.09.04.749349v1.29","doi-asserted-by":"publisher","DOI":"10.1101\/2025.06.16.659792"},{"key":"2026091004100726000_2026.09.04.749349v1.30","doi-asserted-by":"publisher","DOI":"10.1002\/jcb.10645"},{"key":"2026091004100726000_2026.09.04.749349v1.31","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2818.2006.01706.x"},{"key":"2026091004100726000_2026.09.04.749349v1.32","doi-asserted-by":"publisher","DOI":"10.1098\/rsob.240318"},{"key":"2026091004100726000_2026.09.04.749349v1.33","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2024.114720"},{"key":"2026091004100726000_2026.09.04.749349v1.34","doi-asserted-by":"publisher","DOI":"10.1080\/15548627.2020.1739442"},{"key":"2026091004100726000_2026.09.04.749349v1.35","doi-asserted-by":"publisher","DOI":"10.1002\/1873-3468.12979"},{"key":"2026091004100726000_2026.09.04.749349v1.36","doi-asserted-by":"publisher","DOI":"10.1016\/s0021-9258(19)47429-2"},{"key":"2026091004100726000_2026.09.04.749349v1.37","doi-asserted-by":"publisher","DOI":"10.4161\/auto.6845"},{"key":"2026091004100726000_2026.09.04.749349v1.38","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-021-00768-9"},{"key":"2026091004100726000_2026.09.04.749349v1.39","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2020.108468"},{"key":"2026091004100726000_2026.09.04.749349v1.40","unstructured":"World Health Organization. Leishmaniasis. 2023 Jan. Available: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/leishmaniasis"},{"key":"2026091004100726000_2026.09.04.749349v1.41","unstructured":"Cama VA , Kamb Ma . Is cutaneous leishmaniasis autochthonous in the United States? American Society of Tropical Medicine and Hygiene. 2023."},{"key":"2026091004100726000_2026.09.04.749349v1.42","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0000585"},{"key":"2026091004100726000_2026.09.04.749349v1.43","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.adh9902"},{"key":"2026091004100726000_2026.09.04.749349v1.44","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbagen.2011.06.004"},{"key":"2026091004100726000_2026.09.04.749349v1.45","doi-asserted-by":"publisher","DOI":"10.1021\/tx8003906"},{"key":"2026091004100726000_2026.09.04.749349v1.46","doi-asserted-by":"publisher","DOI":"10.1039\/d0ra02934c"},{"key":"2026091004100726000_2026.09.04.749349v1.47","doi-asserted-by":"publisher","DOI":"10.1021\/tx100389r"},{"key":"2026091004100726000_2026.09.04.749349v1.48","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2024.150244"},{"key":"2026091004100726000_2026.09.04.749349v1.49","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2407-7-1"},{"key":"2026091004100726000_2026.09.04.749349v1.50","doi-asserted-by":"publisher","DOI":"10.1111\/imcb.12394"},{"key":"2026091004100726000_2026.09.04.749349v1.51","doi-asserted-by":"publisher","DOI":"10.1007\/s00204-023-03660-8"},{"key":"2026091004100726000_2026.09.04.749349v1.52","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00764-20\/FORMAT\/EPUB"},{"key":"2026091004100726000_2026.09.04.749349v1.53","doi-asserted-by":"publisher","DOI":"10.1016\/j.redox.2026.104071"},{"key":"2026091004100726000_2026.09.04.749349v1.54","doi-asserted-by":"publisher","DOI":"10.1080\/15548627.2018.1474992"},{"key":"2026091004100726000_2026.09.04.749349v1.55","doi-asserted-by":"publisher","DOI":"10.1371\/JOURNAL.PONE.0085262"},{"key":"2026091004100726000_2026.09.04.749349v1.56","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M117.808675"},{"key":"2026091004100726000_2026.09.04.749349v1.57","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2021.748738"},{"key":"2026091004100726000_2026.09.04.749349v1.58","doi-asserted-by":"publisher","DOI":"10.1093\/carcin\/bgn095"},{"key":"2026091004100726000_2026.09.04.749349v1.59","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jpcb.4c04373"},{"key":"2026091004100726000_2026.09.04.749349v1.60","doi-asserted-by":"publisher","DOI":"10.4172\/jcsb.1000026"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.09.04.749349","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T11:10:18Z","timestamp":1789038618000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.09.04.749349"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,7]]},"references-count":60,"URL":"https:\/\/doi.org\/10.64898\/2026.09.04.749349","relation":{},"subject":[],"published":{"date-parts":[[2026,9,7]]},"subtype":"preprint"}}